adsl-1 Antibody
Description
Molecular Characterization
Target: ADSL (Adenylosuccinate Lyase)
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Gene ID: 158 (NCBI)
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UniProt ID: P30566
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Function: Catalyzes two reactions in de novo purine synthesis:
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Structure: 55 kDa protein with enzymatic activity dependent on fumarate production
Antibody Applications
Cancer Biology
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Breast Cancer:
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Prostate Cancer:
Neurodevelopment
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ADSL deficiency causes:
Comparative Antibody Performance
| Supplier | Catalog # | Clonality | Applications |
|---|---|---|---|
| Proteintech | 15264-1-AP | Polyclonal | WB, IHC, IF/ICC, FC |
| Atlas Antibodies | HPA000525 | Polyclonal | WB, ICC, IHC |
| Novus Biologicals | NBP1-87406 | Polyclonal | WB, ICC, IHC |
Technical Considerations
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Q&A
Basic Research Questions
How to validate ADSL-1 antibody specificity in novel model systems?
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Perform siRNA-mediated ADSL knockdown with rescue experiments using siRNA-resistant ADSL constructs to confirm target specificity .
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Include species-specific positive controls (human, mouse, rat) and compare observed molecular weight (55 kDa) against calculated mass (54.9 kDa) .
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Validate using peptide-blocking assays with the immunogen (ADSL fusion protein Ag7332) .
What are optimal storage conditions for long-term antibody stability?
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Store at -20°C in PBS with 50% glycerol and 0.02% sodium azide .
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Avoid repeated freeze-thaw cycles; aliquot into single-use volumes for high-throughput experiments .
How to address discrepancies between observed and predicted molecular weights?
What controls are essential for Western blot/IHC experiments?
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Knockout controls: Use ADSL-depleted cell lysates (e.g., siRNA-treated LNCaP/C4-2B cells) .
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Isotype controls: Rabbit IgG for polyclonal antibody validation .
Advanced Research Questions
How does ADSL depletion impact DNA damage response pathways?
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ADSL silencing increases 53BP1 foci formation (≥5 foci/cell) by 3.8-fold, indicating DNA damage accumulation .
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Combine with ATM inhibitors (e.g., KU5593) to differentiate replication stress from direct DNA damage .
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Monitor RPA2 nuclear intensity as a marker of replication stress .
What methodologies confirm ADSL's role in cell cycle regulation?
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Flow cytometry: ADSL knockdown increases G1-phase cells by 22% (p<0.05) via p53/p21 activation .
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Co-staining: Ki67 (proliferation) with p53 (cell cycle arrest) in ADSL-depleted RPE-1 cells .
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Rescue experiments: Express siRNA-resistant ADSL* to restore Ki67 positivity .
How to investigate ADSL's oncogenic role in prostate cancer?
| Method | Key Parameters |
|---|---|
| siRNA knockdown | Use siRNA 1 (CCAGUUUCCUGCAGCUCUUTT) |
| Xenograft models | Monitor tumor growth in ADSL-silenced C4-2B |
| Pathway analysis | Assess CDK4, CDC2, and Bcl2 expression |
What neurodevelopmental models reveal ADSL deficiency effects?
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Zebrafish embryos: Show neuroectodermal defects with ADSL morpholino knockdown .
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Chicken embryos: Display reduced retinal proliferation (Ki67-negative cells) .
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Human RPE-1 cells: Quantify differentiation markers (vimentin/CK20) post-silencing .
How to resolve conflicting results in ADSL isoform detection?
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Map antibody epitopes using truncated recombinant proteins (e.g., AA 1-310 vs. 185-280) .
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Compare reactivity across species (human vs. dog/guinea pig) using cross-species WB .
Methodological Best Practices
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For co-staining with cell cycle markers, use sequential immunofluorescence with 0.1% Triton-X permeabilization .
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In proliferation assays, combine Trypan Blue exclusion with β-galactosidase staining to distinguish quiescence from senescence .
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For clinical correlations, analyze ADSL expression in TCGA prostate adenocarcinoma datasets alongside cell cycle pathway genes .
